Expression vector delivery system and use thereof for inducing an immune response
Embodiments of a novel system for delivering an expression vector encoding an antigen to a subject that allows for spatiotemporal control over stimulation of the subject's immune response to the antigen are provided. In some embodiments, the expression vector delivery system includes a polymer linked to an adjuvant in prodrug form that can form polymer nanoparticles and enter a cell (such as an immune cell) under physiological conditions. In some embodiments, the adjuvant is linked to the polymer by an enzyme degradable labile bond, the cleavage of which activates the adjuvant to stimulate an immune response.
1. A method of inducing an immune response to an antigen in a subject, comprising:
administering locally to a target location in the subject a therapeutically effective amount of an immunogenic composition comprising a polymer linked to an adjuvant prodrug and an expression vector comprising a nucleic acid molecule encoding the antigen; wherein the polymer linked to the adjuvant prodrug and the expression vector form polymer nanoparticles in the immunogenic composition;
the polymer nanoparticles can enter cells under physiological conditions;
the adjuvant of the adjuvant prodrug is a toll-like receptor 7/8 agonist comprising the structure set forth as:
and
the hydrazine is masked by linkage to a linker connected to the polymer, wherein the linker comprises an enzyme-degradable labile bond, wherein cleavage of the enzyme-degradable labile bond by an intracellular enzyme releases the adjuvant from the polymer to unmask the hydrazine and activates the adjuvant to induce the immune response to the antigen in the subject.
2. The method of claim 1 , wherein the expression vector is a DNA plasmid expression vector that does not comprise any CpG motifs.
3. The method of claim 1 , wherein the polymer is a cationic polymer and the expression vector is linked to the cationic polymer by an electrostatic interaction.
4. The method of claim 3 , wherein the cationic polymer is a poly(ethylenimine) polymer, a poly(lysine) polymer, or a poly(arginine) polymer.
5. The method of claim 1 , wherein the ratio of adjuvant prodrug to monomer of the polymer is from 1:100 to 1:1 mol/mol.
6. The method of claim 1 , wherein the polymer comprises a plurality of monomers comprising from 5 monomers to 500 monomers.
7. The method of claim 1 , wherein the adjuvant prodrug is activated to enhance the immune response from 1-10 days following administration of the immunogenic composition to the subject.
8. The method of claim 7 , wherein the adjuvant prodrug is activated to enhance the immune response from 5-10 days following administration of the immunogenic composition to the subject.
9. The method of claim 1 , wherein the adjuvant prodrug is not activated to enhance the immune response until after the nucleic acid molecule encoding the antigen is expressed in the subject.
10. The method of claim 1 , wherein the enzyme-degradable labile bond is a protease-cleavable labile bond, and wherein cleavage of the protease-cleavable labile bond by an intracellular protease activates the adjuvant to induce the immune response against the antigen in the subject.
11. The method of claim 1 , wherein the intracellular protease is a cathepsin.
12. The method of claim 11 , wherein the linker comprises a cathepsin-cleavable peptide comprising L-amino acids or D-amino acids comprising the amino acid sequence set forth as one of: KPLR (SEQ ID NO: 2), KLRP (SEQ ID NO: 3), SLVR (SEQ ID NO: 4), or SLRV (SEQ ID NO: 5), and cathepsin cleavage of the peptide cleaves the labile bond and activates the adjuvant.
13. The method of claim 1 , wherein the antigen is a tumor associated antigen, a viral antigen, a bacterial antigen, or a protozoan antigen.
14. The method of claim 5 , wherein the ratio of adjuvant prodrug to monomer of the polymer is from 1:20 to 1:10 mol/mol.